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  • Direct On-Chip Differentiation of Intestinal Tubules from Induced Pluripotent Stem Cells.

Direct On-Chip Differentiation of Intestinal Tubules from Induced Pluripotent Stem Cells.

International journal of molecular sciences (2020-07-18)
Elena Naumovska, Germaine Aalderink, Christian Wong Valencia, Kinga Kosim, Arnaud Nicolas, Stephen Brown, Paul Vulto, Kai S Erdmann, Dorota Kurek
ABSTRACT

Intestinal organoids have emerged as the new paradigm for modelling the healthy and diseased intestine with patient-relevant properties. In this study, we show directed differentiation of induced pluripotent stem cells towards intestinal-like phenotype within a microfluidic device. iPSCs are cultured against a gel in microfluidic chips of the OrganoPlate, in which they undergo stepwise differentiation. Cells form a tubular structure, lose their stem cell markers and start expressing mature intestinal markers, including markers for Paneth cells, enterocytes and neuroendocrine cells. Tubes develop barrier properties as confirmed by transepithelial electrical resistance (TEER). Lastly, we show that tubules respond to pro-inflammatory cytokine triggers. The whole procedure for differentiation lasts 14 days, making it an efficient process to make patient-specific organoid tubules. We anticipate the usage of the platform for disease modelling and drug candidate screening.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Fluorescein isothiocyanate–dextran, average mol wt 150,000, (FITC:Glucose = 1:160)
Sigma-Aldrich
[Leu15]-Gastrin I human, ≥95% (HPLC)
Sigma-Aldrich
Tetramethylrhodamine isothiocyanate–Dextran, average mol wt 4,400
Sigma-Aldrich
Nicotinamide, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
N-Acetyl-L-cysteine, BioReagent, suitable for cell culture
Sigma-Aldrich
SB 202190 monohydrochloride hydrate, ≥98% (HPLC)